Autosomal dominant polycystic kidney disease (ADPKD) is a hereditary disorder caused by mutations in the pkd1 and pkd2 genes, leading to the progressive formation of renal cysts. Tolvaptan, a vasopressin 2 receptor (V₂R) antagonist, is the first-line treatment for ADPKD; however, its efficacy is limited and usually associated with significant hepatotoxicity. In this study, we developed a series of tolvaptan derivatives with varying affinities and binding kinetics. Using embryonic kidney cyst model and pkd1 knockout mouse, we screened the V₂R antagonists based on residence time and evaluated their therapeutic effects. We found that the inhibitory effect of these antagonists correlated more with receptor residence time than with affinity, as evidenced by indicators such as the index of cystic area in ex vivo models and the kidney weight index (kidney weight / body weight) in in vivo models. Using isolated kidney perfusion models further demonstrated that prolonged receptor residence time resulted in sustained target occupancy. Our results suggest that the pharmacological effects of V2R antagonists are critically dependent on receptor residence time. This study provides new insights into the development of therapeutic agents for ADPKD.